Effective Generation of Functional Pancreatic β Cells from Human-Derived Dental Stem Cells of Apical Papilla and Bone-Marrow-Derived Stem Cells: A Comparative Study

Abstract

Diabetes Mellitus Type 1 is an autoimmune disease that occurs due to the destruction of insulin-producing cells (β cells), resulting in hyperglycemia. Therefore, diabetic patients depend on insulin treatment for the rest of their lives. Stem cells are considered a promising cellular therapy to replace the nonfunctional beta cells with functional and mature beta cells. Hence, in this study, we aimed to examine the potential of dental stem cells of apical papilla (SCAP) to differentiate into functional islet cell aggregates (ICAs), compared to the ICA generated from bone-marrow-derived stem cells (BM-MSCs). Our strategy was to induce the differentiation of SCAP and BM-MSCs into a definitive endoderm. The success of endodermal differentiation was determined by measuring the expression of definitive endodermal markers, FOXA2 and SOX-17, by flow cytometry. Next, the maturity and functionality of the differentiated cells were evaluated by measuring the amount of insulin and C-peptide secreted by the derived ICAs using ELISA. Additionally, the expression of mature beta cell markers—insulin, C-peptide, glucagon and PDX-1—was detected through confocal microscopy, while the staining of the mature islet-like clusters was detected by using diphenythiocarbazone (DTZ). Our results have shown that both SCAP and BM-MSCs were sequentially committed to a definitive pancreatic endoderm and β-cell-like cells by upregulating the expression of FOXA2 and SOX17 significantly (**** p < 0.0000 and *** p = 0.0001), respectively. Moreover, the identity of ICAs was confirmed by DTZ-positive staining, as well as by the expression of C-peptide, Pdx-1, insulin and glucagon at day 14. It was noted that at day 14, differentiated ICAs released insulin and C-peptides in a significant manner (* p < 0.01, *** p = 0.0001), respectively, exhibiting in vitro functionality. Our results demonstrated for the first time that SCAP could be differentiated into pancreatic cell lineage in a similar manner to BM-MSCs, suggesting a new unambiguous and nonconventional source of stem cells that could be used for stem cell therapy to treat diabetes.

Ameliorating and refining islet organoids to illuminate treatment and pathogenesis of diabetes mellitus

“…Endodermal ASCs, such as pancreatic ductal progenitor cells, PP cells, hepatic stem cells, and gastric stem cells, can directly differentiate into insulin-producing β-cells [ 14 , 27 , 28 ]. Additionally, cells from the mesodermal layer, such as bone marrow mesenchymal stem cells (BMSCs), adipose tissue stem cells (ADSCs), dental pulp stem cells, and ectodermal cells like amniotic epithelial cells, can also be induced to transdifferentiate into β-cells [ 29 , 30 ]. However, the efficiency of differentiating ASCs into β cells varies widely across studies, and their germline origin may be the main reason.…”

Islet Like Cells Induced from Umbilical Cord Mesenchymal Stem Cells with Neonatal Bovine Pancreatic Mesenchymal Exosomes for Treatment of Diabetes Mellitus

“…Cell clusters obtained from the above group D experiment were collected, washed three times with PBS, and stained with diphenyltetrazine (DTZ) [15]. They were transferred to 1.5 ml EP, 0.05 % trypsin EDTA (Life sciences, Beijing) was added at 37°C for 10 min by gently blowing the pipette.…”